Compositions and methods of making polymerizing nucleic acids

Abstract

Provided herein are compositions and methods of making high density nucleic acid polymers.

1 - 63 . (canceled) 
     
     
         64 . A method of making a graft polymer, the method comprising:
 (i) reacting a plurality of oligonucleotide monomers with a polymerization catalyst or initiator, wherein each of said plurality of oligonucleotide monomers comprises a polymerizable monomer covalently bound to an oligonucleotide;   (ii) terminating said reacting with a chain terminator or transfer agent;   wherein the graft polymer has the formula FX1:
   R 1 —[M(O)] n —R 2    (FX1)
 
   wherein,
 n is an integer from 2 to 1000; 
 M is the polymerized product of the polymerizable monomer; 
 O is the oligonucleotide covalently bound to M; 
 R 1  and R 2  are terminal polymer moieties; and 
   wherein 100% of the polymerized polymerizable monomers are each individually attached directly to the oligonucleotide or to the oligonucleotide through a covalent linker.   
     
     
         65 . The method of  claim 64 , wherein the graft polymer is a graft-through polymer. 
     
     
         66 . The method of  claim 64 , wherein the step of reacting comprises radical polymerization, controlled radical polymerization, reversible addition-fragmentation chain transfer (RAFT) polymerization, atom transfer radical polymerization (ATRP), ring-opening metathesis polymerization (ROMP), anionic and cationic polymerizations, free radical living polymerization, acyclic diene metathesis polymerization, radiation-induced polymerization, ring-opening olefin metathesis polymerization, polycondensation reactions, or iniferter-induced polymerization. 
     
     
         67 . The method of  claim 64 , wherein the polymerization catalyst or initiator comprises an ROMP initiator. 
     
     
         68 . The method of  claim 64 , wherein the polymerizable monomer is N-substituted-5-norbornene-2,3-dicarboximide, wherein the substitution comprises the oligonucleotide. 
     
     
         69 . The method of  claim 64 , wherein the oligonucleotide comprises at least 3 nucleobases and at least 2 different nucleobases, at least 5 nucleobases and at least 3 different nucleobases, or at least 10 nucleobases and at least 4 different nucleobases. 
     
     
         70 . The method of  claim 64 , wherein the graft polymer comprises a brush polymer. 
     
     
         71 . The method of  claim 64 , wherein R 1  comprises a substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl. 
     
     
         72 . The method of  claim 64 , wherein wherein R 2  comprises a substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl. 
     
     
         73 . The method of  claim 64 , wherein M(O) is: 
       
         
           
           
               
               
           
         
         wherein,
 L 1  is independently a bond, —O—, —NH—, —COO—, —S—, —SO 2 —, —SO 3 —, —SO 4 —, —SO 2 NH—, —NHC(O)—, —C(O)NH—, —NHC(O)O—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene; and 
 R 4  is the oligonucleotide. 
 
       
     
     
         74 . A method of making an amphiphilic block graft copolymer, the method comprising:
 (i) reacting a plurality of oligonucleotide monomers with a polymerization catalyst thereby forming a hydrophilic block portion, wherein each of said plurality of oligonucleotide monomers comprises a polymerizable monomer covalently bound to an oligonucleotide;   (ii) reacting said hydrophilic block portion with a plurality of hydrophobic monomers and said polymerization catalyst thereby forming said amphiphilic block graft copolymer, wherein each of said plurality of hydrophobic monomers comprises said polymerizable monomer covalently bound to a hydrophobic moiety;   
       wherein the amphiphilic block graft copolymer has the formula (FX2):
   R 1 —[M(O)] n —[M(P)] m —R 2    (FX2)
 
 
       wherein,
 n is an integer from 2 to 1000; 
 m is an integer from 2 to 1000; 
 M is the polymerized product of the polymerizable monomer; 
 O is the oligonucleotide; 
 P is the hydrophobic moiety; and 
 R 1  and R 2  are terminal polymer moieties. 
 
     
     
         75 . The method of  claim 74 , wherein the oligonucleotide comprises at least 3 nucleobases and at least 2 different nucleobases, at least 5 nucleobases and at least 3 different nucleobases, or at least 10 nucleobases and at least 4 different nucleobases. 
     
     
         76 . The method of  claim 74 , wherein the amphiphilic block graft copolymer comprises at least 3 oligonucleotide, at least 5 oligonucleotide branches, or at least 10 oligonucleotide branches. 
     
     
         77 . The method of  claim 74 , wherein the amphiphilic block graft copolymer comprises a linear backbone comprising a polynorbornyl chain. 
     
     
         78 . The method of  claim 74  further comprising atom transfer radical polymerization (ATRP), ring-opening metathesis polymerization (ROMP), anionic and cationic polymerizations, free radical living polymerization, radiation-induced polymerization, ring-opening olefin metathesis polymerization, polycondensation reactions, or iniferter-induced polymerization. 
     
     
         79 . A method of internalizing a nucleic acid into the interior of a cell comprising:
 contacting a cell with a graft polymer or a graft copolymer;   wherein the graft polymer or the graft copolymer has the formula FX1 or FX2:
   R 1 —[M(O)] n —R 2    FX1;
 
   R 1 —[M(O)] n —[M(P)] m —R 2    FX2;
 
   wherein
 n is an integer from 2 to 1000; 
 m is an integer from 2 to 1000; 
 R 1  and R 2  are terminal polymer moieties; 
   
       
         
           
           
               
               
           
         
         
           Olig is independently an oligonucleotide covalently attached directly to M or to M through a covalent linker; 
           HM is independently a hydrophobic moiety covalently attached to M; and 
           R 1  and R 2  are independently terminal polymer moieties. 
         
       
     
     
         80 . The method of  claim 79 , wherein the graft copolymer comprises an amphiphilic graft copolymer. 
     
     
         81 . The method of  claim 79 , further comprising regulating an mRNA level in the cell. 
     
     
         82 . The method of  claim 79 , further comprising detecting a first nucleic acid in the cell, detecting a DNA sequence in the cell, detecting an RNA sequence in the cell, or any combination thereof. 
     
     
         83 . The method of  claim 79 , wherein the graft polymer comprises a second nucleic acid capable of hybridizing the first nucleic acid, a nucleic acid capable of hybridizing the DNA sequence, a nucleic acid capable of hybridizing the RNA sequence, or any combination thereof.